Bacterial Toxin Research Citations

2.2. EAE induction

EAE was induced in female and male mice at the age of 12 weeks as described earlier with some modifications (Chechneva et al., 2011). In brief, mice were injected subcutaneously at two flank sites with an emulsion of myelin oligodendrocyte glycoprotein peptide fragment that spans amino acids 35–55 (MOG) (New England Peptide) in incomplete Freund’s adjuvant supplemented with 500 μg Mycobacterium (M.) tuberculosis H37Ra (Difco). Our previously described EAE model with MOG prepared by sonication causes severe EAE in C57BL/6 mice (Chechneva et al., 2011). Sonication changes biochemical composition of MOG epitopes and/or M. tuberculosis causing an enhanced immune response (Adelmann et al., 1995). In KO mice, sonicated MOG causes mild EAE. EAE in KO and Floxed control mice was induced by MOG (300 μg) emulsion prepared by quick sonication (3 × 5 sec on ice). To induce mild EAE in WT mice, MOG (150 μg) was homogenized with 2 syringes connected by a stopcock (twice 10 min with 3 h interval on ice). Emulsion was kept in the dark at 4 °C between mixing and before preparing into individual doses in syringes. All mice received 200 ng of pertussis toxin (PTX) (List Biological Labs) intraperitoneally (i. p.) on day 0 and 2. …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

… Two hours later, the mice were i.p. injected with Pertussis Toxin (List Biological laboratories, Campbell, CA, USA) freshly prepared in ice-cold PBS (200ng in 100µL …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

EAE induction and exosomes treatment

… To induce EAE, female C57BL/6J mice were immunized with 200 μg MOG35–55 (Sinobioway Co., Ltd., Zhangzhou, China) in complete Freund’s adjuvant (Chondrex Inc., WA, USA) with 4 mg/mL heated-killed M. Tuberculosis (H37Ra, non-viable). Pertussis toxin (List Biological Laboratories Inc., CA, USA) was intraperitoneally injected at 400 ng on days 0 and 2. After 15-20 days, mice displayed the peak of the disease showing complete paralysis of the tail and hind limbs. …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

4.5. Treatment of Stx2 on the Chip

Stx2 (List labs; Campbell, CA, USA) was diluted in the DMEM/F12 as supplemented with 5% (v/v) FBS, 100 unit/mL penicillin, and 100 μg/mL streptomycin. One hundred microliters of Stx2 was added only in the gut module, i.e., after assembly of the gut and kidney modules.

• Product #162L – Shiga Toxin 2 from Escherichia coli

Intramuscular injections

All surgeries were conducted under aseptic conditions on P2 neonates anesthetized by deep hypothermia. For induction, pups were placed in latex sleeves gently buried in crushed ice for 3–5 min and maintenance (up to 15 min) was achieved by placing anesthetized pups on a cold pack (3–4 °C). Following small incisions of the skin to expose the targeted muscles, the viral cocktail (or 0.5% Cholera toxin subunit B (CTB) (List Labs) for labeling of the infrahyoids) was injected via a pneumatic dispense system (Picospritzer) connected to a glass pipette (tip diameter ca. 0.1 mm) mounted on a 3D micromanipulator to guide insertion in the desired muscle. …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt

Methods
… Full-length TcdA and TcdB toxins were purchased from List Biological Laboratories Inc. …

• Product #152C – Toxin A from Clostridium difficile

• Product #155 – Toxin B from Clostridium difficile

Stromal Cell Culture

Mouse lung fibroblasts were isolated from whole lung explants by digestion … For experiments, fibroblasts were used between passages 4 and 6. Fibroblasts were serum-starved with DMEM/F-12 medium containing 1% FBS for 24 hr before stimulation with IFN-β and lipopolysaccharide (LPS). Confluent cells were stimulated with LPS from Escherichia coli serotype 0111: B4 (lot no. 2014641; List Biological Laboratories, Campbell, CA) at 100 ng/ml and IFN-β (PBL Interferon Source; Piscataway, NJ) at 100 units/ml. After 4 hr of stimulation, the medium was removed, cells were washed with PBS, and RNA was extracted using RNAeasy Plus Mini Kit (Qiagen).

• Product #201 – LPS from Escherichia coli O111:B4

… Pertussis toxin was purchased from List Biological Laboratories, INC. (Campbell, CA, USA) …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

2.4.1. Retrograde labeling

… A pulled glass pipette (~20 μm outer tip diameter) was attached to the arm of the stereotaxic apparatus. A solution of 0.25% CTB (List Biological Labs, Campbell, CA, USA) in sodium phosphate buffer (pH 7.5) was backfilled through the pipette tip using negative pressure, then a wire connected to a current source (Stoelting) was inserted into the tracer solution. …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt

… For the induction of conventional EAE, naïve female C57BL/6 mice, 8-12 weeks old, were injected subcutaneously (s.c.) in their flanks with 0.1ml of an emulsion containing 100µg of MOG35-55 and 400µg heat-killed M. tuberculosis (H37Ra, Difco) in Incomplete Freunds adjuvant (IFA, Difco) and, the same day and 2 days later, received intravenously (i.v.) 200ng of B. pertussis Toxin (PTX) (List Laboratories) …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)